Preparing and identifying a monoclonal antibody against human colorectal carcinoma

陈曦; 黄悦; 于彬; 梅峰; 韩鹃; 周德山

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Preparing and identifying a monoclonal antibody against human colorectal carcinoma

更新时间：2026-01-09

- Preparing and identifying a monoclonal antibody against human colorectal carcinoma
- Journal of Army Medical University Vol. 29, Issue 10, Pages: 896-898(2007)
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  CLC： R392-33;R392.2;R735
- Published：2007
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陈曦,黄悦,于彬,梅峰,韩鹃,周德山.结直肠癌单克隆抗体的制备及生物学特性鉴定[J].陆军军医大学学报(原第三军医大学学报),2007,29(10):896-898.

　CHEN Xi,HUANG Yue,YU Bin,MEI Feng,HAN Juan,ZHOU De-shan.Preparing and identifying a monoclonal antibody against human colorectal carcinoma[J].J Amry Med Univ (J Third Mil Med Univ),2007,29(10):896-898.
陈曦,黄悦,于彬,梅峰,韩鹃,周德山.结直肠癌单克隆抗体的制备及生物学特性鉴定[J].陆军军医大学学报(原第三军医大学学报),2007,29(10):896-898. DOI：

　CHEN Xi,HUANG Yue,YU Bin,MEI Feng,HAN Juan,ZHOU De-shan.Preparing and identifying a monoclonal antibody against human colorectal carcinoma[J].J Amry Med Univ (J Third Mil Med Univ),2007,29(10):896-898. DOI：

摘要

目的制备结直肠癌特异性单克隆抗体，为深入研究相关肿瘤的发病机制提供新的实验工具。方法用人结直肠癌患者的手术标本制备细胞悬液免疫小鼠，按常规方法行细胞融合，经免疫组织化学染色筛选阳性克隆，并对其抗体的特性和生物学性状等进行分析。结果共获得32个阳性克隆，其中针对结直肠癌较特异性的阳性克隆有2个(克隆株2-3

15-8)。免疫组化染色显示，克隆株2-3培养上清和腹水与结直肠癌组织的冰冻切片反应的阳性率可达100%。癌旁正常组织的阳性率较低，且染色强度远弱于癌组织，当抗体稀释度超过1∶6 000时反应阴性，而肿瘤组织仍呈强阳性。从结肠癌的分化程度来看，分化较差的肿瘤组织，包括中、低分化的癌组织染色较强。另外，免疫染色阳性产物主要位于肿瘤的腺腔和正常黏膜表面，提示该抗体所识别的抗原可能为结直肠癌细胞和黏膜上皮分泌的黏蛋白等抗原。结论本研究成功地制备了两株针对结直肠癌的较特异的单克隆抗体，可作为实验研究相关蛋白在结直肠癌及其他肿瘤发病中的作用机制的方法。

Abstract

Objective To produce monoclonal antibodies against colorectal carcinoma and provide a new tool for study of the mechanisms in related cancers. Methods Specimens of colorectal carcinoma were freshly obtained from surgery. Balb/c mice were immunized with cell suspension from colorectal carcinoma tissues. Fusion was done as classical hybridoma method. Positive hybridomas were selected by immunohistochemical staining. The nature and biological characteristic of the selected monoclonal antibodies was investigated. Results Thirty-two positive hybridomas were selected. Two (2-3

15-8) of these hybridomas secreted antibodies binding specifically to human colorectal carcinoma. By immunohistochemical staining

all of colorectal carcinoma cases was positive for 2-3 antibody. In contrast

normal colon and rectal mucosa showed much weaker staining

and was totally negative when 2-3 ascites antibody was diluted over 1∶6 000. In addition

2-3 antibody reacted strongly with moderately differentiated and poorly differentiated colorectal carcinoma. The immunoreactive substance was observed on luminal contents of colorectal carcinoma tubules and on apical surface of normal epithelium

indicating that the antigen recognized by 2-3 antibody may be a kind of mucin secreted by colorectal carcinoma and normal epithelium and may participate in tumor invasion and metastasis. Conclusion We successfully produced two monoclonal antibodies against colorectal carcinoma.

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